Leaf samples of Anemia villosa (Anemiaceae) were collected in four distinct places in Rio de Janeiro State, Brazil. Two coastal land populations are located in Itacoatiara and Imbuhy rocky outcrops, municipality of Niterói, and two populations in the inland mountain region of the state, Pedra Dubois, municipality of Santa Maria Madalena, and in Lumiar, municipality of Nova Friburgo. Five leaves of each one of five randomly chosen individuals were collected in each site. The four populations analyzed were very similar in anatomic qualitative characteristics. ANOVA results for the quantitative variables showed significant differences between all populations. The PCA separated the populations analyzed in two groups: one formed by the coastal land and other formed by the inland mountain.